Rapid unbiased isolation and in situ RNA analysis of circulating tumor cells using a magnetic micropore-based diagnostic chip

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ABSTRACT: Pancreatic cancer is the fourth most common cause of cancer related death in the United States, with a five year survival rate of only 4%. We have recently shown that circulating pancreatic cells (CPCs) can be detected in the blood at the onset of the disease cycle in both mice and humans. These findings present an opportunity to develop a non-invasive diagnostic for pancreatic cancer that has enormous potential to improve patient outcomes. The current gold-standard for circulating tumor cell (CTC) detection fails to measure the extremely sparse and heterogeneous CPCs. Even next generation microfluidic technologies, which do not rely on epithelial markers, have limited utility due to the inherently low-throughput of microfluidics and the large sample volumes of blood (V > 10 mL) necessary for ultra-rare cell detection. To address these challenges, we are developing a new approach to rare cell detection, using magnetic micropores, which combines the benefits of micro-scale sorting with extremely fast flow rates (100x faster than typical microfluidic approaches2,5-7). In addition to its improved throughput, our technique is robust against unprocessed clinical samples, allowing rare CPCs and CPC clusters to be isolated directly from whole blood. Moreover, we are integrating this approach with ultra-rapid on-chip RNA fluorescence in-situ hybridization (RNA FISH), enabling single molecule, multiplexed RNA analysis in individual rare cells. Our device, which combines the above features into a self-contained, automated format, is designed to extract the RNA expression of single CPCs in clinical settings.